The generation and properties of human cortical organoids as a disease model for malformations of cortical development

As three-dimensional“organ-like”aggregates,human cortical organoids have emerged as powerful models for studying human brain evolution and brain disorders with unique advantages of humanspecificity,fidelity and manipulation.Human cortical organoids derived from human pluripotent stem cells can elaborately replicate many of the key properties of human cortical development at the molecular,cellular,structural,and functional levels,including the anatomy,functional neural network,and interaction among different brain regions,thus facilitating the discovery of brain development and evolution.In addition to studying the neuro-electrophysiological features of brain cortex development,human cortical organoids have been widely used to mimic the pathophysiological features of cortical-related disease,especially in mimicking malformations of cortical development,thus revealing pathological mechanism and identifying effective drugs.In this review,we provide an overview of the generation of human cortical organoids and the properties of recapitulated cortical development and further outline their applications in modeling malformations of cortical development including pathological phenotype,underlying mechanisms and rescue strategies.

Cellular interplay to 3D in vitro microphysiological disease model:cell patterning microbiota-gut-brain axis

The microbiota-gut-brain axis(MGBA)has emerged as a key prospect in the bidirectional communication between two major organ systems:the brain and the gut.Homeostasis between the two organ systems allows the body to function without disease,whereas dysbiosis has long-standing evidence of etiopathological conditions.The most common communication paths are the microbial release of metabolites,soluble neurotransmitters,and immune cells.However,each pathway is intertwined with a complex one.With the emergence of in vitro models and the popularity of three-dimensional(3D)cultures and Transwells,engineering has become easier for the scientific understanding of neurodegenerative diseases.This paper briefly retraces the possible communication pathways between the gut microbiome and the brain.It further elaborates on three major diseases:autism spectrum disorder,Parkinson’s disease,and Alzheimer’s disease,which are prevalent in children and the elderly.These diseases also decrease patients’quality of life.Hence,understanding them more deeply with respect to current advances in in vitro modeling is crucial for understanding the diseases.Remodeling of MGBA in the laboratory uses many molecular technologies and biomaterial advances.Spheroids and organoids provide a more realistic picture of the cell and tissue structure than monolayers.Combining them with the Transwell system offers the advantage of compartmentalizing the two systems(apical and basal)while allowing physical and chemical cues between them.Cutting-edge technologies,such as bioprinting and microfluidic chips,might be the future of in vitro modeling,as they provide dynamicity.

Human pluripotent stem cell-derived kidney organoids:Current progress and challenges

Human pluripotent stem cell(hPSC)-derived kidney organoids share similarities with the fetal kidney.However,the current hPSC-derived kidney organoids have some limitations,including the inability to perform nephrogenesis and lack of a corticomedullary definition,uniform vascular system,and coordinated exit path-way for urinary filtrate.Therefore,further studies are required to produce hPSC-derived kidney organoids that accurately mimic human kidneys to facilitate research on kidney development,regeneration,disease modeling,and drug screening.In this review,we discussed recent advances in the generation of hPSC-derived kidney organoids,how these organoids contribute to the understanding of human kidney development and research in disease modeling.Additionally,the limitations,future research focus,and applications of hPSC-derived kidney organoids were highlighted.

Bioengineered skin organoids:from development to applications

Signifcant advancements have been made in recent years in the development of highly sophisticated skin organoids.Serving as three-dimensional(3D)models that mimic human skin,these organoids have evolved into complex structures and are increasingly recognized as efective alternatives to traditional culture models and human skin due to their ability to overcome the limitations of two-dimensional(2D)systems and ethical concerns.The inherent plasticity of skin organoids allows for their construction into physiological and pathological models,enabling the study of skin development and dynamic changes.This review provides an overview of the pivotal work in the progression from 3D layered epidermis to cyst-like skin organoids with appendages.Furthermore,it highlights the latest advancements in organoid construction facilitated by state-of-the-art engineering techniques,such as 3D printing and microfuidic devices.The review also summarizes and discusses the diverse applications of skin organoids in developmental biology,disease modelling,regenerative medicine,and personalized medicine,while considering their prospects and limitations.

Success rate of current human-derived gastric cancer organoids establishment and influencing factors:A systematic review and meta-analysis

BACKGROUND Human-derived gastric cancer organoids(GCOs)are widely used in gastric cancer research;however,the culture success rate is generally low.AIM To explore the potential influencing factors,and the literature on successful culture rates of GCOs was reviewed using meta-analysis.METHODS PubMed,Web of Science,and EMBASE were searched for studies.Two trained researchers selected the studies and extracted data.STATA 17.0 software was used for meta-analysis of the incidence of each outcome event.The adjusted Methodological Index for Non-Randomized Studies scale was used to assess the quality of the included studies.Funnel plots and Egger’s test were used to detect publication bias.Subgroup analyses were conducted for sex,tissue source,histo-logical classification,and the pathological tumor-node-metastasis(pTNM)cancer staging system.RESULTS Eight studies with a pooled success rate of 66.6%were included.GCOs derived from women and men had success rates of 67%and 46.7%,respectively.GCOs from surgery or biopsy/endoscopic submucosal dissection showed success rates of 70.9%and 53.7%,respectively.GCOs of poorly-differentiated,moderately-differentiated and signet-ring cell cancer showed success rates of 64.6%,31%,and 32.7%,respectively.GCOs with pTNM stages I-II and III-IV showed success rates of 38.3%and 65.2%,respectively.Y-27632 and non-Y-27632 use showed success rates of 58.2%and 70%,respectively.GCOs generated with collagenase were more successful than those constructed with Liberase TH and TrypLE(72.1%vs 71%,respectively).EDTA digestion showed a 50%lower success rate than other methods(P=0.04).CONCLUSION GCO establishment rate is low and varies by sex,tissue source,histological type,and pTNM stage.Omitting Y-27632,and using Liberase TH,TrypLE,or collagenase yields greater success than EDTA.

In vitro engineered models of neurodegenerative diseases

Neurodegeneration is a catastrophic process that develops progressive damage leading to functional andstructural loss of the cells of the nervous system and is among the biggest unavoidable problems of our age.Animalmodels do not reflect the pathophysiology observed in humans due to distinct differences between the neuralpathways,gene expression patterns,neuronal plasticity,and other disease-related mechanisms in animals andhumans.Classical in vitro cell culture models are also not sufficient for pre-clinical drug testing in reflecting thecomplex pathophysiology of neurodegenerative diseases.Today,modern,engineered techniques are applied to developmulticellular,intricate in vitro models and to create the closest microenvironment simulating biological,biochemical,and mechanical characteristics of the in vivo degenerating tissue.In THIS review,the capabilities and shortcomings ofscaffold-based and scaffold-free techniques,organoids,and microfluidic models that best reflect neurodegeneration invitro in the biomimetic framework are discussed.

Visualization analysis of research hotspots and trends on gastrointestinal tumor organoids

BACKGROUND Gastrointestinal tumor organoids serve as an effective model for simulating cancer in vitro and have been applied in basic biology and preclinical research.Despite over a decade of development and increasing research achievements in this field,a systematic and comprehensive analysis of the research hotspots and future trends is lacking.AIM To address this problem by employing bibliometric tools to explore the publication years,countries/regions,institutions,journals,authors,keywords,and references in this field.METHODS The literature was collected from Web of Science databases.CiteSpace-6.2R4,a widely used bibliometric analysis software package,was used for institutional analysis and reference burst analysis.VOSviewer 1.6.19 was used for journal cocitation analysis,author co-authorship and co-citation analysis.The‘online platform for bibliometric analysis(https://bibliometric.com/app)’was used to assess the total number of publications and the cooperation relationships between countries.Finally,we employed the bibliometric R software package(version R.4.3.1)in R-studio,for a comprehensive scientific analysis of the literature.RESULTS Our analysis included a total of 1466 publications,revealing a significant yearly increase in articles on the study of gastrointestinal tumor organoids.The United States(n=393)and Helmholtz Association(n=93)have emerged as the leading countries and institutions,respectively,in this field,with Hans Clevers and Toshiro Sato being the most contributing authors.The most influential journal in this field is Gastroenterology.The most impactful reference is"Long term expansion of epithelial organs from human colon,adenoma,adenocarcinoma,and Barrett's epithelium".Keywords analysis and citation burst analysis indicate that precision medicine,disease modeling,drug development and screening,and regenerative medicine are the most cutting-edge directions.These focal points were further detailed based on the literature.CONCLUSION This bibliometric study offers an objective and quantitative analysis of the research in this field,which can be considered as an important guide for next scientific research.

Application of patient-derived tumor models in anticancer drug development and individualized medicine

Malignant tumor is the second leading cause of death due to its high incidence, lack of effective treatment and poor prognosis. The evaluation of anticancer drugs used to based on NCI-60 cell line models, but the limited heterogeneity and the divorce from clinical practice of models lead to extremely low success rate of novel anticancer drugs during clinical trials (less than 10%). In recent years, because of the high heterogeneity and human derived tumor matrix, patient-derived tumor models have been gradually applied to the preclinical evaluation of various antitumor drugs, which shows certain advantages in predicting the clinical efficacy of antitumor drugs. Optimize the drug combination through patient-derived tumor models to achieve individualized medicine has gradually become an indispensable strategy in clinical cancer therapy. The current review summarized the development of patient-derived tumor models, characterized the application, advantages and challenges of them in preclinical antitumor drug evaluation and clinical precise medicine, which will provide a scientific basis and novel insights for further basic research, drug development and clinical application.

CMIP 6 models simulation of the connection between North/South Pacific Meridional Mode and ENSO

The subtropical North and South Pacific Meridional Modes(NPMM and SPMM)are well known precursors of El Niño-Southern Oscillation(ENSO).However,relationship between them is not constant.In the early 1980,the relationship experienced an interdecadal transition.Changes in this connection can be attributed mainly to the phase change of the Pacific decadal oscillation(PDO).During the positive phase of PDO,a shallower thermocline in the central Pacific is responsible for the stronger trade wind charging(TWC)mechanism,which leads to a stronger equatorial subsurface temperature evolution.This dynamic process strengthens the connection between NPMM and ENSO.Associated with the negative phase of PDO,a shallower thermocline over southeastern Pacific allows an enhanced wind-evaporation-SST(WES)feedback,strengthening the connection between SPMM and ENSO.Using 35 Coupled Model Intercomparison Project Phase 6(CMIP6)models,we examined the NPMM/SPMM performance and its connection with ENSO in the historical runs.The great majority of CMIP6 models can reproduce the pattern of NPMM and SPMM well,but they reveal discrepant ENSO and NPMM/SPMM relationship.The intermodal uncertainty for the connection of NPMM-ENSO is due to different TWC mechanism.A stronger TWC mechanism will enhance NPMM forcing.For SPMM,few models can simulate a good relationship with ENSO.The intermodel spread in the relationship of SPMM and ENSO owing to SST bias in the southeastern Pacific,as WES feedback is stronger when the southeastern Pacific is warmer.

Hepatitis B virus infection modeling using multi-cellular organoids derived from human induced pluripotent stem cells

Chronic infection with hepatitis B virus(HBV)remains a global health concern despite the availability of vaccines.To date,the development of effective treatments has been severely hampered by the lack of reliable,reproducible,and scalable in vitro modeling systems that precisely recapitulate the virus life cycle and represent virus-host interactions.With the progressive understanding of liver organogenesis mechanisms,the development of human induced pluripotent stem cell(iPSC)-derived hepatic sources and stromal cellular compositions provides novel strategies for personalized modeling and treatment of liver disease.Further,advancements in three-dimensional culture of self-organized liver-like organoids considerably promote in vitro modeling of intact human liver tissue,in terms of both hepatic function and other physiological characteristics.Combined with our experiences in the investigation of HBV infections using liver organoids,we have summarized the advances in modeling reported thus far and discussed the limitations and ongoing challenges in the application of liver organoids,particularly those with multi-cellular components derived from human iPSCs.This review provides general guidelines for establishing clinical-grade iPSC-derived multi-cellular organoids in modeling personalized hepatitis virus infection and other liver diseases,as well as drug testing and transplantation therapy.

Organoids: a novel modality in disease modeling

Limitations of monolayer culture conditions have motivated scientists to explore new models that can recapitulate the architecture and function of human organs more accurately.Recent advances in the improvement of protocols have resulted in establishing three-dimensional(3D)organ-like architectures called‘organoids’that can display the characteristics of their corresponding real organs,including morphological features,functional activities,and personalized responses to specific pathogens.We discuss different organoid-based 3D models herein,which are classified based on their original germinal layer.Studies of organoids simulating the complexity of real tissues could provide novel platforms and opportunities for generating practical knowledge along with preclinical studies,including drug screening,toxicology,and molecular pathophysiology of diseases.This paper also outlines the key challenges,advantages,and prospects of current organoid systems.

Brain organoids are new tool for drug screening of neurological diseases

At the level of in vitro drug screening,the development of a phenotypic analysis system with highcontent screening at the core provides a strong platform to support high-throughput drug screening.There are few systematic reports on brain organoids,as a new three-dimensional in vitro model,in terms of model stability,key phenotypic fingerprint,and drug screening schemes,and particula rly rega rding the development of screening strategies for massive numbers of traditional Chinese medicine monomers.This paper reviews the development of brain organoids and the advantages of brain organoids over induced neurons or cells in simulated diseases.The paper also highlights the prospects from model stability,induction criteria of brain organoids,and the screening schemes of brain organoids based on the characteristics of brain organoids and the application and development of a high-content screening system.

Modernising autism spectrum disorder model engineering and treatment via CRISPR-Cas9:A gene reprogramming approach

A neurological abnormality called autism spectrum disorder(ASD)affects how a person perceives and interacts with others,leading to social interaction and communication issues.Limited and recurring behavioural patterns are another feature of the illness.Multiple mutations throughout development are the source of the neurodevelopmental disorder autism.However,a well-established model and perfect treatment for this spectrum disease has not been discovered.The rising era of the clustered regularly interspaced palindromic repeats(CRISPR)-associated protein 9(Cas9)system can streamline the complexity underlying the pathogenesis of ASD.The CRISPR-Cas9 system is a powerful genetic engineering tool used to edit the genome at the targeted site in a precise manner.The major hurdle in studying ASD is the lack of appropriate animal models presenting the complex symptoms of ASD.Therefore,CRISPR-Cas9 is being used worldwide to mimic the ASD-like pathology in various systems like in vitro cell lines,in vitro 3D organoid models and in vivo animal models.Apart from being used in establishing ASD models,CRISPR-Cas9 can also be used to treat the complexities of ASD.The aim of this review was to summarize and critically analyse the CRISPRCas9-mediated discoveries in the field of ASD.

Unraveling pathological mechanisms in neurological disorders:the impact of cell-based and organoid models

Cell-based models are a promising tool in deciphering the molecular mechanisms underlying the pathogenesis of neurological disorders as well as aiding in the discovery and development of future drug therapies.The greatest challenge is creating cell-based models that encapsulate the vast phenotypic presentations as well as the underlying genotypic etiology of these conditions.In this article,we discuss the recent advancements in cell-based models for understanding the pathophysiology of neurological disorders.We reviewed studies discussing the progression of cell-based models to the advancement of three-dimensional models and organoids that provide a more accurate model of the pathophysiology of neurological disorders in vivo.The better we understand how to create more precise models of the neurological system,the sooner we will be able to create patient-specific models and large libraries of these neurological disorders.While three-dimensional models can be used to discover the linking factors to connect the varying phenotypes,such models will also help to understand the early pathophysiology of these neurological disorders and how they are affected by their environment.The three-dimensional cell models will allow us to create more specific treatments and uncover potentially preventative measures in neurological disorders such as autism spectrum disorder,Parkinson’s disease,Alzheimer’s disease,and amyotrophic lateral sclerosis.

头颈部恶性肿瘤研究模型的演化

恶性肿瘤发生发展的机制探索以及抗癌药物治疗疗效的评估均有赖于各种体内与体外研究模型的建立。近几十年间,随着生物医学技术的快速发展,恶性肿瘤的体内外研究模型也发生了巨大的变化。基因检测技术从单基因到多基因的进展促进了生物信息学飞速发展和恶性肿瘤概念的转变;体外细胞研究模型从单层的二维培养、原代培养向立体的三维构型发展,从而更好地重现肿瘤组织的细胞间交互作用与功能;体内动物研究模型由传统的致癌物诱导、细胞或组织形成移植瘤逐渐演变为基因编辑的动物模型或人源性肿瘤异种移植模型,从而可以针对性地研究相关基因在肿瘤发生发展中的作用;传统的临床研究也从简单的临床回顾性研究更多地向前瞻性研究转变,Ⅰ期/Ⅱ期/Ⅲ期临床研究,研究者发起的临床研究以及真实世界临床研究,这些研究为临床研究增添了活力。目前恶性肿瘤研究模型存在的主要不足包括模型的单一性、对肿瘤微环境的模拟不足、动物肿瘤模型与人类肿瘤差异性,以及缺乏对个性化医疗的考量。未来仍需要进一步研发和优化研究模型,并更有效地将不同模型整合起来,形成一个优化的整体实验模型系统。本文将系统回顾恶性肿瘤研究模型的演化并对相关模型进行阐述,为科研工作者进行恶性肿瘤的研究提供合理的研究模型。

器官芯片——更具前景的体外模型

传统的体外模型具有不可避免的局限性,与人体试验相比,在评估药物疗效和不良反应方面存在显著差异。器官芯片技术在生理环境和功能芯片上模拟人体器官,具有高保真的生理或病理生理功能,为药物开发提供了巨大的创新前景。本文主要从体内各系统角度介绍器官芯片的研究进展和应用,同时提出目前器官芯片发展过程中存在的局限性和未来的发展方向。

高级别浆液性卵巢癌类器官模型的建立及鉴定

目的 建立并鉴定高级别浆液性卵巢癌类器官模型。方法 选取行卵巢癌细胞减灭术的10例高级别浆液性卵巢癌患者的卵巢癌组织,将卵巢癌组织消化成单细胞,在基质胶中培养7天,构建高级别浆液性卵巢癌类器官模型。比较类器官与原发肿瘤的形态学特征、免疫组化特征和苏木精-伊红(HE)染色结果。结果 10例患者的高级别浆液性卵巢癌类器官模型均成功构建。光学显微镜显示类器官数量可观,形态呈类圆形3D立体结构。HE染色显示,在高倍镜下,类器官细胞核异型性明显,局部区域形成微乳头状结构。免疫组化显示黏蛋白16(MUC16)强阳性,p53弥漫性强阳性,类器官与原发肿瘤的免疫组化染色情况一致。结论 体外成功构建的高级别浆液性卵巢癌类器官模型具有明显的上皮性特征,与原发肿瘤高度相似,可用于卵巢癌的药物筛选和基础研究建模。

肝器官芯片在生物医学研究中的应用进展

肝脏具有复杂结构和多种功能,包括血糖调控、蛋白合成、解毒和药物代谢等,在维持人体正常生理活动中起着重要作用。传统的二维细胞培养和动物模型已被广泛用于肝脏生理或疾病研究,但它们在反映人体组织真实微环境和对药物反应等方面仍存在一定局限。因此,建立高仿真度肝脏体外模型对于肝病研究、药效与毒性评价至关重要。本文概述了传统肝脏体外模型在实现近生理复杂微环境模拟、肝组织特异性功能准确复现等方面的局限性,总结了以器官芯片为代表的新型肝脏体外模型的设计策略、技术特点及其在生物医学领域的研究进展。文中重点介绍了肝器官芯片仿生构筑和实现肝组织微环境模拟的关键要素,包括多细胞组分、肝窦/肝小叶结构、生化因子梯度和流体因素等,并对未来结合其他先进手段(如类器官、生物材料和基因编辑等)等,建立高度生理相关性的肝器官芯片和微生理系统的发展前景予以展望。

脑类器官在再生医学中的研究进展

脑类器官是一种基于人多能干细胞的三维体外模型,能够模拟人脑的细胞异质性、结构和功能。再生医学是一个多学科交叉的领域,致力于应用工程学和生物学手段修复因年龄、疾病或外伤而受损的组织或器官。脑类器官技术作为再生医学领域的一种重要手段,具有广阔的应用前景和重要的科学意义。近年来,利用组织工程和诱导因子分化技术,研究人员成功构建出不同脑区的脑类器官模型,可用于模拟脑损伤或修复病变组织。本文将系统介绍包括大脑皮层、海马、纹状体、中脑、丘脑及下丘脑、小脑和视网膜在内的脑区特异类器官构建技术的最新进展,总结其在再生医学领域中的应用,并概括当前脑类器官应用面临的挑战,如异质性大、缺乏脉管系统和成熟度较低等。这将加深对人类大脑的理解,并增强脑类器官在基础研究和临床研究中的进一步应用。

糖尿病肾病模型:动物模型、二维细胞模拟及三维类器官模型

背景:近几年来,人类多能干细胞衍生的肾类器官及糖尿病肾病啮齿动物模型取得了一定的进展。然而,由于糖尿病肾病发病受环境因素及遗传因素共同作用,发病机制复杂,并且对于此类患者的临床治疗方法需要因人而异。因此,需要开发更灵活和综合的方法,从而发现强有力的临床前证据,以支持对糖尿病肾病患者进行更有针对性的干预。目的:从动物模型、二维细胞培养模拟及三维类器官模型等方面综述了糖尿病肾病模型的研究进展,为进一步的研究提供线索和思路。方法:以“糖尿病,糖尿病肾病,糖尿病肾病模型,糖尿病肾病动物模型,类器官,肾脏类器官,糖尿病肾病细胞模型,糖尿病肾病病证结合动物模型”为中文检索词,“Diabetes,diabetic nephropathy,diabetic kidney diseases,diabetic nephropathy models,diabetic nephropathy animal models,organoids,diabetic and organoids,diabetic and kidney organoids,kidney organoids,diabetic nephropathy cell models,diabetic nephropathy syndrome combined animal models”为英文检索词,检索中国知网及PubMed数据库,最终纳入101篇文献进行综述分析。结果与结论:糖尿病肾病体内模型和体外模型是深入研究糖尿病肾病发病机制的有力工具。动物模型可以观察到多系统之间的相互作用;二维细胞培养操作简便且成本较低;新兴的肾类器官填补了二维和整体水平之间的空白,无种属差异且一定程度模拟了人肾脏的复杂性。随着类器官技术的不断发展,肾类器官有望为探索糖尿病肾病的发病机制、病理生理过程和药物筛选提供一个新的视角。